title: No human virus sequences detected by next-generation sequencing in benign verrucous skin tumors occurring in BRAF-inhibitor-treated patients.
abstract: Patients treated with BRAF inhibitors (e.g. vemurafenib), a novel targeted therapy for advanced melanoma harbouring certain BRAF mutations, develop numerous adverse cutaneous side effects, including skin tumors such as squamous cell carcinoma or non-malignant verruciform keratinocyte proliferations, termed 'BRAF-inhibitor-associated verrucous keratosis (BAVK) lesions'. These keratinocyte proliferations are believed to be caused by paradoxical hyperactivation of the MAPK pathway in cells with wild-type BRAF, but mutated RAS. However, due to the clinical and histological verruca-like appearance of these lesions, additional aetiologic cofactors, such as infectious agents (i.e. oncogenic viruses), might be suspected. Therefore, we performed 454 high-throughput sequencing of BAVK lesions from vemurafenib-treated patients on the transcript level to identify actively transcribed viral sequences of known [e.g. human papilloma viruses (HPV)] or even yet-unknown viruses. Next-generation sequencing did not identify transcripts of any human viruses out of 1 595 161 reads obtained from BAVK lesions of four patients. Nevertheless, all controls were recognized correctly, and the detection of sequences derived from the cutaneous microbiome (e.g. skin commensals and bacterial phages) confirmed the validity and sensitivity of the sequencing data. Our results are consistent with preliminary histological and immunohistochemical findings recently reported by others, who also failed to detect the expression of HPV proteins in BAVK. Although the patient number is limited and we cannot exclude the possibility of having missed a viral transcript of very low abundance, our study argues against a viral aetiology of BRAF-inhibitor-associated verruciform keratoses occurring under vemurafenib.
authors.affiliation: Institute of Virology, Hannover Medical School, Hannover, Germany.
authors.forename: Tina
authors.initials: T
authors.lastname: Ganzenmueller
authors.forename: Elias
authors.initials: E
authors.lastname: Hage
authors.forename: Yuri
authors.initials: Y
authors.lastname: Yakushko
authors.forename: Jeanette
authors.initials: J
authors.lastname: Kluba
authors.forename: Sabrina
authors.initials: S
authors.lastname: Woltemate
authors.forename: Vivien
authors.initials: V
authors.lastname: Schacht
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Schulz
authors.forename: Ralf
authors.initials: R
authors.lastname: Gutzmer
mesh_terms: D000328:Adult; D000368:Aged; D001706:Biopsy; D018289:Carcinoma, Verrucous / Q000821:virology*; D049109:Cell Proliferation; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007211:Indoles / Q000627:therapeutic use; D015603:Keratinocytes / Q000166:cytology; D008297:Male; D008545:Melanoma / Q000188:drug therapy*; D008875:Middle Aged; D009154:Mutation; D027383:Papillomaviridae / Q000235:genetics; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D012878:Skin Neoplasms / Q000150:complications / Q000188:drug therapy* / Q000821:virology*; D013449:Sulfonamides / Q000627:therapeutic use; D000077484:Vemurafenib
chemical_list: D004279:DNA, Viral; D007211:Indoles; D013449:Sulfonamides; D000077484:Vemurafenib; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf
keywords: BRAF inhibitors; cutaneous side effects; next-generation sequencing; verrucous keratosis; virus